Low dose oral beta-lactamase protects the gut microbiome from oral beta-lactam-mediated damage in dogs

Sheila Connelly; Brian Fanelli; Nur A. Hasan; Rita R. Colwell; Michael Kaleko; Sheila Connelly; Brian Fanelli; Nur A. Hasan; Rita R. Colwell; Michael Kaleko

doi:10.3934/publichealth.2019.4.477

AIMS Public Health

2019, Volume 6, Issue 4: 477-487. doi: 10.3934/publichealth.2019.4.477

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Research article

Low dose oral beta-lactamase protects the gut microbiome from oral beta-lactam-mediated damage in dogs

1.
Synthetic Biologics, Inc., Rockville, MD, United States
2.
CosmosID, Inc., Rockville, MD, United States
3.
University of Maryland Institute for Advanced Computer Studies, College Park, MD, United States

Received: 05 September 2019 Accepted: 05 November 2019 Published: 12 November 2019

Antibiotics, while lifesaving, damage the gut microbiome and can precipitate proliferation of pathobionts. A strategy to preserve gut microbiome integrity is to eliminate biologically active antimicrobials excreted into the gastrointestinal tract (GI) without negatively affecting antibiotic therapeutic efficacy. Clinical proof of concept was achieved with SYN-004 (ribaxamase), a beta-lactamase enzyme formulated for oral delivery with intravenous penicillins and cephalosporins. Ribaxamase inactivated intestinal ceftriaxone, protected the gut microbiome, and significantly reduced the incidence of Clostridioides difficile disease. For use with oral beta-lactam antibiotics, a delayed release formulation of ribaxamase, SYN-007, was engineered for dissolution in the lower small intestine distal to the site of oral antibiotic absorption. In dogs that received oral amoxicillin, SYN-007 reduced microbiome disruption without interfering with amoxicillin systemic absorption. Here, a study to determine the lowest effective dose of SYN-007 was performed. Dogs received amoxicillin (40 mg/kg, PO, TID) +/– SYN-007 (PO, TID) at three doses, 10 mg, 3 mg, or 1 mg for five days. Serum amoxicillin levels, measured after the first and last antibiotic doses, were not significantly different +/– SYN-007 at all dose levels indicating that SYN-007 did not interfere with amoxicillin systemic absorption. Microbiome analyses demonstrated that amoxicillin significantly reduced bacteria richness and microbiome diversity resulting in altered microbiome composition. However, with all doses of SYN-007, microbiome richness and diversity were not significantly different from pretreatment and changes in microbiome composition were attenuated. These data demonstrate that effective SYN-007 doses can be reduced at least 10-fold while maintaining gut microbiome preservation. The potential to employ low SYN-007 doses to protect the gut microbiota has important implications for enhancing therapeutic outcomes for patients receiving oral beta-lactam antibiotics while simultaneously reducing cost per dose and ultimately, healthcare expenses.
- antibiotic,
- gut microbiome,
- beta-lactam,
- beta-lactamase
Citation: Sheila Connelly, Brian Fanelli, Nur A. Hasan, Rita R. Colwell, Michael Kaleko. Low dose oral beta-lactamase protects the gut microbiome from oral beta-lactam-mediated damage in dogs[J]. AIMS Public Health, 2019, 6(4): 477-487. doi: 10.3934/publichealth.2019.4.477

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Abstract

Antibiotics, while lifesaving, damage the gut microbiome and can precipitate proliferation of pathobionts. A strategy to preserve gut microbiome integrity is to eliminate biologically active antimicrobials excreted into the gastrointestinal tract (GI) without negatively affecting antibiotic therapeutic efficacy. Clinical proof of concept was achieved with SYN-004 (ribaxamase), a beta-lactamase enzyme formulated for oral delivery with intravenous penicillins and cephalosporins. Ribaxamase inactivated intestinal ceftriaxone, protected the gut microbiome, and significantly reduced the incidence of Clostridioides difficile disease. For use with oral beta-lactam antibiotics, a delayed release formulation of ribaxamase, SYN-007, was engineered for dissolution in the lower small intestine distal to the site of oral antibiotic absorption. In dogs that received oral amoxicillin, SYN-007 reduced microbiome disruption without interfering with amoxicillin systemic absorption. Here, a study to determine the lowest effective dose of SYN-007 was performed. Dogs received amoxicillin (40 mg/kg, PO, TID) +/– SYN-007 (PO, TID) at three doses, 10 mg, 3 mg, or 1 mg for five days. Serum amoxicillin levels, measured after the first and last antibiotic doses, were not significantly different +/– SYN-007 at all dose levels indicating that SYN-007 did not interfere with amoxicillin systemic absorption. Microbiome analyses demonstrated that amoxicillin significantly reduced bacteria richness and microbiome diversity resulting in altered microbiome composition. However, with all doses of SYN-007, microbiome richness and diversity were not significantly different from pretreatment and changes in microbiome composition were attenuated. These data demonstrate that effective SYN-007 doses can be reduced at least 10-fold while maintaining gut microbiome preservation. The potential to employ low SYN-007 doses to protect the gut microbiota has important implications for enhancing therapeutic outcomes for patients receiving oral beta-lactam antibiotics while simultaneously reducing cost per dose and ultimately, healthcare expenses.

Acknowledgments

This work was supported by Synthetic Biologics, Inc. The authors acknowledge Cristina Freire and Ting Chen at Aptuit, an Evotec Company, formerly Kuecept, Ltd, for production of SYN-007 formulated capsules, and Steven D. Sloneker, Study Director at Calvert Laboratories, Inc. for providing outstanding support with dog dosing and sample collections. We thank Dr. Christian Furlan-Freguia for critical review of the manuscript, and Leslie Marlow and Dr. Stephen Altieri for disclosure reviews of the manuscript.

Ethics Approval

All animal procedures were approved by and conducted in accordance with principles and guidelines established by the Calvert Institutional Animal Care and Use Committee in accordance with the Animal Welfare Act at Calvert Laboratories, Inc. (Scott, PA). Calvert Laboratories, Inc. is fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC).

Authors' contributions

SC and MK designed the experimental studies and interpreted the data. BF and NAH coordinated the sequencing of the fecal DNA and performed the sequencing data analyses. SC and BF produced the figures. BF submitted the fecal DNA metagenomics sequencing data to the SRA. SC wrote the manuscript. BF, NAH, RRC, and MK reviewed and edited the manuscript. All authors read and approved the final manuscript.

Conflict of interests

The authors declare the following potential conflicts of interest with respect to the research, authorship and/or publication of this article: SC and MK are employees of Synthetic Biologics, Inc. RRC is the founder of CosmosID, Inc., a fee-for-service provider engaged by Synthetic Biologics, Inc. BF and NAH are employees of CosmosID, Inc.

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